The transcription factor Cbfa1 regulates osteoblast differentiation and expression of genes necessary for the development of a mineralized phenotype. The purpose of this study was to determine if Cbfa1 and BSPII gene expression are influenced by implant surface microtopography. Osteoblasts were cultured on 600-grit (grooved) or sandblasted (roughened) cpTi implant discs. Mineralization was evaluated by Alizarin-Red-S staining. Real Time PCR was used for quantitative analysis of Cbfa1 and BSPII gene expression. Enhanced mineralization was seen in osteoblasts grown on roughened implant surfaces relative to tissue culture plastic. Real Time PCR showed significant (P < 0.05) increases in Cbfa1 gene expression in cells grown on roughened, as compared with grooved, implant surfaces. BSPII gene expression was also increased on rough surfaces in the UMR cells, but was reduced in the rat calvarial osteoblast cultures. These results suggest that osteoblast gene expression and mineralization are affected by roughened implant surface microtopographies during osseointegration of dental implants.
To mimic skeletal muscle tissues in vitro, native
and transgenic spider silk/silkworm silks were seeded with C2C12 myoblasts
to observe if these three-dimensional substrates are preferable to
a traditional two-dimensional polystyrene cell culture surface. Silks
were wound around an acrylic chassis to produce a novel, three-dimensional
cell culture device with suspended muscle fibers that genetically
and morphologically resemble native skeletal muscle tissue. The transgenic
spider silk/silkworm silk has never before been studied for this application.
Genetic expression verified skeletal muscle lineage and differentiation,
while fluorescent imaging verified contractile protein synthesis.
Genetic analysis also revealed an increase in expression of the Myh2 contractile protein gene on silkworm silks, particularly
on the transgenic silk. Mechanical properties and protein secondary
structure content of the silks indicated correlation between substrate
properties and Myh2 gene expression. This increase
in contractile protein gene expression suggests that biologically
derived silk substrates that are suspended may be a preferable substrate
for in vitro muscle modeling because of the proteinaceous
character and mechanical flexibility of the silk.
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